Method of preparing and using isoflavones for the treatment of blood related illnesses

ABSTRACT

A composition is prepared by extracting phytochemicals from plant matter and is administered to provide treatment for cardiovascular medical conditions, such as: excessive bloodstream cholesterol, the risk of heart disease, abnormal blood lipid profiles, and abnormal vascular effects. This composition is enriched preferably with two or more fractions of plant matter, namely: isoflavones, lignans, saponins, sapogenins, catechins and phenolic acids. The isoflavones are selected from a group including malonyl, acetyl, glucoside and aglycone. Soy is the preferred source of these chemicals; however, other plants may also be used, such as wheat, psyllium, rice, oats, red clover, kudzu, alfalfa, flax, and cocoa. The composition is a dietary supplement in a concentrated form, preferably in an easy to consume form, for treatment of various cardiovascular conditions and various other related disorders.

This is a division of Ser. No. 09/162,038, filed Sep. 28, 1998 (a formalapplication which replaced provisional application Ser. No. 60/060,549filed Oct. 2, 1997) now U.S. Pat. No. 6,261,565, which in turn, is acontinuation-in-part of Ser. No. 09/035,588, filed Mar. 5, 1998, nowU.S. Pat. No. 6,033,714, which in turn, is a continuation-in-part ofSer. No. 08/868,629, filed Jun. 4, 1997, now U.S. Pat. No. 5,792,503,which in turn, is a division of Ser. No. 08/614,545, filed Mar. 13,1996, now U.S. Pat. No. 5,702,752.

This invention relates to compositions extracted from vegetable matterand more particularly to phytochemicals, including saponogenins andsaponins, catechins, lignans, phenolic acids, and isoflavones, andespecially those extracted from a family of plants including soy, flax,tea, and cocoa and methods of using these compositions as nutritionalsupplements or food additives.

BACKGROUND

As used herein, the term “isoflavone” includes malonyl, acetyl,glucoside, and aglycone forms of the isoflavones.

Currently there is great interest in treating blood related illnessessuch as excessive levels of cholesterol, coronary disease, abnormalblood lipid profiles and vascular effects. It is thought that at leastsome of these illnesses may be either caused or exacerbated by foods,especially those of an animal origin. Therefore, it is thought that manyof these illnesses are either preventable or treatable by a use ofphytochemicals, especially isoflavones, as a source of supplementalhormones.

It is also though that there are superior results when a plurality ofsuch phytochemicals are consumed in combinations which are tailored toparticular symptoms. A proper diet should contain the desiredphytochemicals. However, a trouble is that many people do not have or donot like the proper kind of diet which provides the desirable effects.Hence, the problem is to furnish the necessary food values in some otherform. Therefore, there is a need for a refining process which willenable a selection of specific phytochemical food values in combinationsthat are desirable for specific blood related illnesses.

Plant materials are known to contain a number of classes of organic lowmolecular weight compounds which exert bioactivity in various animals.Historically, these compounds have been considered to be somewhatnon-nutritive, however, recent scientific evidence now suggests thesecompounds may play an important role in the maintenance of health, inchemoprevention, and in the mitigation of certain conditions or diseasesassociated with the circulation of sex hormones, including sleepdisorders and vaginal dryness.

Edible plants normally contained in the diet, or materials used asherbal remedies/dietary supplements, may contain collections ofstructurally related compounds. These related substances are oftenunique in their amounts and distribution when compared among variousplant sources. The most notable groups of compounds exhibitingbioactivity are known as flavonoids, isoflavones, saponins, lignans,alkaloids, catechins and phenolic acids.

Epidemiology studies relating diet to disease suggest that dietarycomponents may predispose populations to reduced risk of certaindiseases. Far eastern populations consuming soy have reduced rates ofbreast, prostate and colon cancers and coronary heart disease, whilepopulations in Finland have reduced rates of prostate cancer.Researchers are just now studying the specific compounds in the diet tounderstand the basis for the epidemiological observations.

Among the various plants consumed in the diet, several are rich sourcesof phytochemicals. Soy products contain high amounts of isoflavones andsaponins. Unrefined diet grains include plants such as wheat, psyllium,rice, flax and oats that contain lignans. Cocoa contains catechins andphenolic acids. Certain non-dietary plants are also sources of the samechemical molecules, such as lignans and isoflavones in kudzu root or redclovers. Isoflavones and lignans act as weak estrogenic substances. Teaplants are also a rich source of phytochemicals, including catechins andphenolic acids.

Isoflavones can be used alone to treat or prevent breast cancer,prostate cancer, skin cancer, and colon cancer or as mechanisminhibitors. Isoflavones alone may also reduce or prevent varioussymptoms related to the onset and duration of menopause, including hotflashes and osteoporosis. Isoflavones alone may also be effective incertain cardiovascular applications, including heart disease, reducingcholesterol-lipid levels, modulating angiogenesis, and other vasculareffects. Moreover, isoflavones along have been implicated in reducingheadaches, dementia, inflammation, and alcohol abuse, as well asimmunomodulation.

Lignans along have been implicated in preventing or treating breastcancer, prostate cancer and colon cancer as well as reducing hotflashes, preventing osteoporosis and showing antiviral potential.Lignans also have antimitotic and fungicidal activity. A plant lignan,the catecholic nordihydro-guaiaretci acid, was a potent antioxidant onceused by the food industry.

Saponins alone have been implicated in preventing or treating skincancer, colon cancer, reducing serum cholesterol, and inimmunomodulation and antiviral activity. Saponins also exhibitantioxidant effects and act as free radical scavengers.

Phenolic acids have shown antioxidant activity.

People who eat a high soy diet show reduction of many of theseabove-discussed symptoms. This suggests that ingesting a combination ofthese phytochemicals in a ratio such as that found in soy may result inan additive or synergistic effect. However, a high soy diet has someundesirable effects, including flatulence, undesirable taste, andhesitancy among Western consumers to change their lifestyle toincorporate soy in their diets, even for such benefits.

Isoflavones, which are heterocyclic phenols, are understood to includethe soy compounds genistin, daidzin and glycitein, as well as biochaninA, equol, formononetin, and o-desmethylangolensin and naturalderivatives thereof. These compounds and their aglycone or de-methylatedaglycone forms, such as genistein and daidzein, are believed to havesimilar activities once they are ingested. They are sometimes referredto as phytoestrogens.

Lignans are defined to be compounds possessing a 2,3-dibenzylbutanestructure. They include matairesinol, secoisolariciresinol,lariciresinol, isolariciresinol, nordihydroguaiaretic acid, pinoresinol,olivil, other compounds which may be precursors of enterolactone andenterodiol and modifications thereof, including diglucosides.

Phenolic acids include p-hydrobenzoic acid, protocatechuic acid, andvanillic acid. Other phenolic acids are chlorogenic acid, caffeic acid,ferulic acid, gallic acid, sinapic acid, syringic acid, coumaric acid,cinnamic acid, gentisic acid, salicylic acid, hydroxy benzoic acid andhydroxy phenyl acetic acids and derivatives. This list of phenolic acidsshould be understood to include the various isomers and derivativesfound in the natural vegetable source.

Catechins, or flavan-3-ols, include epigallocatechin, catechin,epicatechin and gallocatechin.

Saponogenins are C-27 sterols in which the side cahin has undergonemetabolic changes to produce a spiroketal. Saponogenins occur naturallyas saponins, which are 3-O-glycosides of the parent steroid ortriterpenes. Digitonin from Digitalis is a saponin. Saponins includeglucosides of sapogenin such as triterpenoids or steroids andsaccharides such as glucose, arabinose, galactose or glucuronic acid.Typical examples of leguminous saponins are glycyrrhizin (glycyrrhetinicacid+glucuronic acid) contained in Glycyrrhiza glahra, soysaponincontained in soybean and alfalfasaponin contained in Medicago sativa.Saponins also include chemical entities identified as triterpene phenolssuch as tomatine, soyasapogenols A, B, C, D, E and F, ginsengosidefraction 3 and 4, medicagenic acid, hederagenin, glycyrrhizin digitonin,quillaja saponin, lucernic acid and zahnic acid. The naturalmodifications of these compounds found in the vegetable source are alsoincluded in this identification.

A need exists for an improved composition consisting substantially ofisoflavones, lignans, saponogenins, saponins, and/or phenolic acidswhich will produce improved results over any of these taken alone.Furthermore, a need exists for a composition in which the beneficialphytochemicals are enriched as compared to their original source. Thispermits individuals to conveniently consume such phytochemicals as anutritional supplement or as a food additive.

SUMMARY OF THE INVENTION

An object of this invention is to provide a convenient way forindividuals to consume isoflavones, lignans, saponins, catechins and/orphenolic acids, either as a nutritional supplement or as an ingredientin a more traditional type of food.

An other object of this invention is to provide an optimized extractcomposition of phytochemicals which is in sufficient concentration to bedelivered in an easy to consume dosasge such as a pill, tablet, capsule,liquid or ingredient in a food including health bars.

Yet another object of this invention is to prepare the phytochemicalextract to be delivered as a topical application in a cream or lotion.In this form, the isoflavones, lignans, saponins, catechins and/orphenolic acids are dispersed and suspended in a suitable liquid or gelmatrix to render a stable cream or lotion as the delivery vehicle.

A further object of this invention is to provide an extract concentratewhich is closely similar in chemical composition to the chemicalentities found in the natural plant source.

In keeping with this aspect of the invention, the isoflavones, lignans,saponins, catechins and/or phenolic acids are extracted from a suitablevegetable source to render a composition which is substantially moreconcentrated than the original material and by more than 5 times in oneor more of the desired bioactive components.

This extract may be used alone or combined with one or more other plantextracts to produce the optimized composition. Further, this extractcomposition may be formulated with one or more other dietary nutrients,such as vitamins, minerals, amino acids, etc., to provide a nutritionalsupplement further optimized for a desired health effect. All theseingredients may be combined with necessary binders, excipients,preservatives, colors and the like known to those in the industry inorder to produce a suitable tablet, capsule, pill, liquid, cream, powderor food ingredient including health bars.

These phytochemicals may be packaged and provided in final form by meansknown to the supplements and food ingredient industries. The materialsare intended to provide health and well-being benefits.

DETAILED DESCRIPTION OF THE INVENTION

The improved composition is obtained by fractionating a plant sourcehigh in isoflavones, lignans and other phytochemicals such as defattedsoybean flakes, soy molasses, soy whey, red clover, alfalfa, flax,cocoa, tea, or kudzu root. These may be fractionated along or incombination with these other plants known to be high in the variousisoflavones, lignans, saponins, catechins and phenolic acids. Thefractionation results in substantially removing water, carbohydrates,proteins, and lipids from the source material. The fractionation methodmay be preferably that disclosed in U.S. Pat. No. 5,702,752 or U.S. Pat.No. 4,428,876, or an extraction using ethyl acetate or n-butanol may beused. U.S. Pat. Nos. 5,702,752 6,017,556; 6,033,714 are assigned to theassignee of this invention.

Other extraction processes, which may be used alone or in combination,include differential solubility, distillation, solvent extraction,adsorptive means, differential molecular filtration and precipitation.

The preferred composition is an improvement over known commercialmaterials regarding the amount of phytochemicals per gram of substanceand the amounts of different phytochemicals present which affectphysiologic function.

These natural substances have been consumed in food sources for longperiods of time and more closely relate to the substances consumed whichprovide the basis for the epidemiological evidence for health benefits.Additional benefits may be derived from improved physical propertiesrelative to phytochemicals chemically modified from their original foodsource form.

The resulting composition is expected to comprise in a preferred form:between 5% and 95% isoflavones, between 0% and 70% lignans, and between2% and 70% saponins and sapogenins. In a more preferred form, thecomposition will be extracted from soy. In another preferred form, thecomposition will contain a ratio of (saponins plus saponogenins) toisoflavones from 1:100 to 100:1, with the isoflavones consistingpredominantly of naturally occurring derivatives of genistein and/or itsprecursor biochanin A and daidzein and/or its precursor formononetin,with a ratio of the genistein derivatives to daidzein derivatives from100:1 to 1:100. Preferably, the isoflavones are predominantlyglycosylated derivatives.

The composition's ratios may be readily varied by changing the plantsource or by combining several plant sources for extraction. Thus, asfurther study shows which phytochemical combinations are moreefficacious for certain health effects, the particular composition willalso vary.

It is known that isoflavones, lignans, and saponins can be usedadvantageously to treat or prevent various cancers, including breastcancer, prostate cancer, skin cancer, and colon cancer.

It is believed that the improved composition will provide increasedbenefits in the form of chemoprevention. Recent experiments appear toconfirm this belief.

EXAMPLE 1

An initial series of animal studies was made to investigate the effectsof dietary soy products on the growth of s.c. (SUBCUTANEOUS) implantedLNCaP in male SCID mice. A high isoflavone-containing soy proteinisolate (SPI) (2.0 mg isoflavones/g SPI) is provided by ProteinTechnology International (St. Louis, Mo.). A soy phytochemicals extract,soy phytochemicals concentrate (SPC) which contains 28.5% total soyisoflavones and a diverse amount of other soy phytochemicals, isprovided by Archer Daniels Midland Company (Decatur, Ill.). Thesematerials were used to prepare six experimental diets. Table 1 showsingredients of the diets.

Eight-week-old male SCID mice were s.c. injected on the right flank with2×10⁶ LNCaP cells from hosts, randomized into six groups (n=10) andassigned to one of the experimental diets. Food intake, body weight, andtumor volume were measured. At the termination of the experiment, bloodsamples were collected and serum separated for PSA analysis. An aliquotof tumor tissues was formalin-fixed, paraffin-embedded, and cut into 4μm sections for in situ histochemical detection of apoptotic cells, andimmunohistochemical analyses of angiogenesis and proliferation. Anotheraliquot was prepared for cell lysates for western blot to determine theexpression of apoptosis-related gene products.

Table 2 summaries the effects of treatment of food intake, body weight,isoflavone intake and tumor volume. Soy products did not significantlyalter food intake or body weight. Compared to casein-fed controls, tumorvolumes from mice treated with SPI (20%), SPC (1.0%), and SPI and SPC(1.0%) were reduced by 12%, 28% (P<0.04), or 40% (P<0.005),respectively. Factorial analysis indicated that there was no significanteffect of protein source on tumor growth. Linear regression analysisindicated that tumor volumes were inversely correlated to total dietaryisoflavones (Tumor volume (cm³)=−0.008+2.121×Isoflavones (mg), R²=0.76,p<0.03).

Table 3 shows the effects of SPC at 1.0% of the diet on apoptosis,proliferation, and angiogenesis of tumors from a pilot study. Itindicates that dietary supplementation of soy phytochemicals inhibitsthe growth of LNCaP tumor in vivo by enhancing apoptosis and inhibitingproliferation of tumor cells. It inhibitory effect on tumor angiogenesisis not significant which may be due to small sample size (n=2).

Results from in vitro study showed that genistein and soy phytochemicalconcentrate inhibited secretion of PSA by LNCaP cells into media. PSAconcentrations were reduced 68% and 74% by 25 and 50 uM of genisteintreatment respectively, and 31% and 42% by 25 and 50 μM of soyphytochemical concentrate treatment respectively.

TABLE 1 uz,8/25 Ingredients of experimental diets (grams) Diet 3 Diet 4Diet 5 Diet 6 Diet 1 Diet 2 Casein/ SPI/ Casein/ SPI/ casein SPI LSPCLSPC HSPC HSP SPI 0 200 0 200 0 200 Casein 200 0 200 0 200 0DL-methionine 3 3 3 3 3 3 Corn starch 150 150 150 150 150 150 Sucrose500 500 500 500 500 500 Cellulose, 50 50 50 50 50 50 BW200 Corn oil 5050 50 50 50 50 Mineral Mix, 35 35 35 35 35 35 S10001¹ Vitamin Mix, 10 1010 10 10 10 V10001¹ Choline 2 2 2 2 2 2 Bitartrate Soy 0 0 2 2 10 10phytochemicals Total (g) 1000 1000 1002 1002 1010 1010 (isoflavones, 0245 341 586 705 950 mg/kg diet) ¹AIN formulation with minor modificationby DR. E. A. Ulman, Research Diets, Inc.

TABLE 2 Final body weight, total food intake. total isoflavone intake,and tumor volume Tumor Food intake Total volume Treatment Body weightgrams/m isoflavone (cm³) Casein 22.4 ± 0.5¹ 46.6 ± 3.1  0.00 ± 0.00 2.32± 0.31² SPI 23.1 ± 0.7 46.2 ± 2.8 17.00 ± 6.37 2.06 ± 0.32 Casein/LSPC21.4 ± 0.7 41.2 ± 3.4 14.03 ± .14 1.88 ± 0.35 SPI/LSPC 22.6 ± 0.6 50.1 ±4.7 29.36 ± 2.76 1.66 ± 0.29* Casein/HSPC 22.2 ± 0.7 44.8 ± 6.1 76.38 ±1.64 ± 0.22* 10.40 SPI/HSPC 22.0 ± 0.6 47.5 ± 1.7 92.53 ± 3.22 1.39 ±0.30** ¹Values are means ± S.E. There are no significant differences offood intake or body weight among treatment groups. ²Compared withcontrol group, SPI/LSPC, casein/HSPC, and SPI/HSPC had significantlysmall tumor volumes *p > 0.04, **p > 0.005

TABLE 3 Effects of treatment on apoptotic index (AI, % TUNEL),proliferation index (PI, % PCNA Staining) and angiogenesis (microvesseldensity) Microvessel Treatment AI (% TUNEL) PI (% PCNA) Density Control(n = 2)  6.07 ± 0.88 60.1 ± 1.1 12.5 ± 3.8 Casein/HSPC (n = 2) 10.75 ±0.54 51.7 ± 1.3  9.7 ± 0.7 P value <0.02 <0.01 >0.05 Values are means ±S.E.

In summary, preliminary results indicate that soy products inhibit thes.c. growth of LNCaP tumor in SCID mice, possibly via induction ofapoptosis, and inhibition of angiogenesis and proliferation.

Isoflavones or lignans can alleviate menopausal-related symptoms such ashot flashes and osteoporosis as well as alleviate symptoms associatedwith menstruation. This is further believed to be due to theirestrogenic activity. It is believed that the improved compositiondescribed here will alleviate these symptoms even more effectively.

Also, isoflavones positively affect various cardiovascular-relatedconditions, including heart disease, cholesterol (saponins alsopositively effect cholesterol), angiogenesis and other vascular effects.It is believed that the improved composition will produce results forthese cardiovascular conditions at least as beneficial as those hithertoknown and at a reduced cost.

As explained earlier, isoflavones, lignans, and saponins are known toindividually positively affect various neurological and immunologicalsymptoms. It is believed that the improved composition will result inalleviating neurological and immunological symptoms at least as well asthose compounds hitherto known and at a reduced cost. Moreover, it wouldbe expected that some synergism would arise out of the combinationdescribed herein.

The improved composition may be administered orally, parenterally, forinstance, subcutaneously, intravenously, intramuscularly,intraperitoneally, by intranasal instillation or by application of anaerosol spray to mucous membranes, or to the skin by an ointment or acream.

Administering the improved composition may be done with any suitablecarrier, in solid or liquid dosage form such as tablets, capsules,powders, soft gels, solutions, suspensions, emulsions, ointments, orcreams. The improved composition may also be administered as a foodsupplement or as a food ingredient.

The amount of the improved composition administered will vary dependingon the person, the mode of administration, and the desired result. Aneffective amount is expected to be 10 mg to 2000 mg/per dose.

EXAMPLE 2 Tablet Manufacture

The composition provided for in this patent may be used to preparetablets or other dosage forms. An example of a capsule preparation isprovided in Example 2. The higher the concentration of the activecomponent, the easier it is to form a tablet or emulsion. This leads toan added ability to incorporate other dietary nutrients. An examplewould be to prepare a phytochemical tablet which incorporates calciumand vitamin E as a supplement to maintain bone health and/or reduce postmenopausal symptoms such as hot flashes. In an example of thisembodiment, a 600 mg dry compression tablet was prepared containing atotal of 125 mg of isoflavones concentrate (50 mg isoflavone compound).Included in the tablet formulation was a source of calcium andmagnesium.

Dry compression tablets were produced by first blending the followingingredients: 4 kg of the improved composition (39.83% isoflavones), 1.91kg sorbitol, 0.095 kb magnesium stearate, and 13.11 kg dicalciumphosphate in a 120 quart capacity Hobart mixer. This blend ofingredients was then dry compressed at 1 ton pressure with a Stokes BB2simple press into tablets having a total weight of 600 mg containing125.53 mg of the improved composition and therefore 50 mg of totalisoflavones.

Alternatively, a phytochemical concentrate may be provided in a singledosage form, a skin cream or as a food ingredient added to conventionalfood in amounts from 10 mg to 2000 mg/per dose, the purpose of which isto exert a positive effect on health and well being. These benefitsinclude: cancer prevention, estrogen and sex hormone related maladies,inhibition of the pituitary-thyroid-gonadotrophic axis, alcoholdependency reduction, modulation of the cardiovascular, immune andnervous systems, antiviral effects and analgesic effects.

EXAMPLE 3

Two-piece gelatin capsules were produced by filling the receiving end ofthe empty size “0” capsules with 0.106 g of the improved composition(44.35% isoflavones) and closed with the capping end, providing acapsule containing 47.2 mg of total isoflavones.

EXAMPLE 4

A comparison between various sources of phytochemical preparations isgiven in Table 4. It is readily seen that the phytochemical componentsof the compositions of the “Isoflavone Concentrate” of this invention issubstantially higher than the corresponding amounts in the naturalvegetable materials. Notably, the amount of glycone isoflavones andsaponins are over 100 times more concentrated compared to the foodsource and over twenty times more concentrated compared to the germ ofthe plant which naturally concentrates these phytochemicals. Comparisonof the “Isoflavone Concentrate” of this invention to other concentratesshows that the isoflavone fraction predominates in these latterproducts, reducing the amount of other healthful phytochemicals.Additionally, the extraction methods of these other products employtechniques which modify the components, particularly the isoflavones, sothat they are not identical to the substances found in the naturalvegetable material (U.S. Pat. No. 5,637,562).

One version of the improved composition was compared to other previouslydescribed compositions. The results are shown in Table 4

TABLE 4 Comparative Products to the Invention Isoflavone IsoflavoneGlycosides in Aglycones in Genistein/ Phenolic Product Product ProductDaidzein Lignans Saponins Acids Example (mg/g) (mg/g) Ratio (mg/g)(mg/g) (mg/g) Improved 401.0 3.37 1.06 to 1 0.2 460.7 25.47 compositionSoybean 1.748-2.776^(a) 0.044^(a)-0.075 1.59-2.7 NA 0.9-3.2^(b) SoyFlour 1.969^(a) 0.045^(a) 3.58 0.0013 2.870^(c) (defatted Soy germ24.32^(d) 0.85^(d) NA 16.7-1.98^(b) NA Product^(c) NA 2.5-6.5^(c)0.5-3.5 NA NA NA patent (PTI) Product^(i) NA 5.1-14.7^(i) 0.433-3.48 NANA NA patent (PTI) Product^(g) NA 1.7-3.5^(g) 0.66-2.86 NA NA NA patent(PTI) PTI NA 970 12.8 NA NA NA product^(h) PTI NA 640 2.0 NA NA NAproduct^(h) Soy Molasses 27.6 0.1 1.37 NA NA 5.788 (dried) Novogen^(i)0.0 550 1-1.7 to 1 NA NA NA ^(a)Wang H. and Murphy P. A., J. Agric. FoodChem 1994, 42, 1666-1673. ^(b)Anderson R. L. and Wolf W. J., J. Nutr125:581S-588S, 1995 ^(c)Seo A. and Morr C. V., J. Agric. Food Chem 1984,32, 530-533. ^(d)Soy Life ™ promotional literature ^(e)WO 95/10530,PCT/US94/10697 ^(f)WO 95/10512, PCT/US94/10699 ^(g)WO 95/10529,PCT/US94/10696 ^(h)NCI paper ^(i)Novogen promotional literature

EXAMPLE 5

The improved composition, containing the glycoside forms of isoflavones,has as one aspect an improved solubility at body temperature over thepreviously described compositions containing the aglycoside forms.

Separate solutions (0.02% in distilled water) were made for genistein,genistin, daidzein, daidzin, and isoflavone concentrate in volumetricflasks. Samples were then placed in a 37° C. water bath for 17 hours,followed by rapid filtration through a 0.2 micron syringe-type filter toremove particulates. Filtered samples were then analyzed for isoflavoneconcentration by HPLC. Results are tabulated as shown in Table 5.

TABLE 5 Differential Solubility of Isoflavone Glycosides vs. AglyconesIsoflavone Genistein Genistin sample (ppm) (ppm) Daidzein (ppm) Daidzin(ppm) Genistein 7.42 Genistin 33.89 Daidzein 3.64 Daidzin 48.51Isoflavone 0.492 30.075 0.672 37.69 Concentrate

The glycoside forms, genistin, and daidzin, are at least 4.57 and 13.32fold higher in concentration at 37° C. than their corresponding aglyconeforms, respectively.

The modifications made to the isoflavones are to remove the carbohydrateattached to the isoflavone moiety. This modification renders theisoflavone less soluble in water. Maintenance of the naturalmodification, glycosylation, enhances solubility. This fact is shown inthe comparative solubility chart of Table 5. This chart shows that thegenistin isoflavone is 4.6 times higher than and the daidzin isoflavoneis 13.3 times higher than the corresponding non-glycosylated form.Higher solubility can lead to better bioavailability to intestinalorganisms. The glycosylation does not inhibit absorption in the gutbecause the intestinal microflora convert the glycone form to theaglycone form before absorption occurs.

EXAMPLE 6 Extraction of Lignans from Flax

Lignans can be readily extracted from flax using this following method.

978 g of defatted flax metal (F1) was extracted with 2000 g of 85%ethanol at 40° C. for 10 minutes, forming a slurry. The resulting slurrywas filtered and extraction was repeated twice with a total of 6000 g ofethanol.

The ethanolic fraction was then evaporated under vacuum at 70° C.,resulting in an aqueous fraction of 1186 g. The aqueous fraction wascombined with 1000 g of water and mixed.

The mixed sample was then ultra-filtered through a 5000 molecular weightcutoff membrane, resulting in a 767 g permeate fraction and a retentatefraction of 1283 g.

The retentate fraction was freeze-dried, resulting in a 27.84 g sample(F2).

The 767 g permeate fraction at 50° C. was fed to a 35 ml bed volume,XAD-4 resin column at a rate of 10 ml/min. The column effluent wascollected and dried, resulting in a 14.8 g sample (F3). XAD-4 is atrademark for an absorbent resin, available from Rohm & Haas.

The column was then eluted with four bed volumes (140 ml) of 70% ethanolat 50° C. The eluted sample was evaporated under vacuum at 70° C. anddried, resulting in a 1.79 g sample (F4). The four fractions were thenanalyzed for their lignan content, measured as the concentration byweight of secoisolariciresinol. As Table 6 shows, this extraction methodenriches lignan concentration.

TABLE 6 LIGNAN CONCENTRATIONS AS SECOISOLARICIRESINOL FRACTION F1 F2 F3F4 SECO. CONC. (mg/g) 2.3 1.9 4.8 13.4 PHENOLIC ACID

While the present invention has been disclosed in terms of the preferredembodiment in order to facilitate a better understanding of theinvention, it should be appreciated that the invention can be embodiedin various ways without departing from the principles of the invention.Therefore, the invention should be understood to include all possibleembodiments, modifications, and equivalents to the described embodimentwhich do not depart form the principles of the invention as set out inthe appended claims.

What is claimed is:
 1. A composition for treatment of cardiovascularconditions, said composition being made from a plant matter in which thecomposition is a medicament enriched in at least two of thephytochemicals selected from the group consisting of isoflavones,lignans, saponins, sapogenins, catechins and phenolic acids, saidmedicament having a therapeutic treatment amount of phytochemicalsselected on a basis of a therapeutic treatment for a cardiovascularcondition.
 2. The composition of claim 1 in which the ratio ofisoflavones to saponins is selected from the range of about 1:10 toabout 10:1.
 3. The composition of claim 1 in which the isoflavones arepresent in an amount from approximately 5% to approximately 90% byweight.
 4. The composition of claim 1 in which said medicament is in aform of a product for oral delivery, said product form being selectedfrom a group consisting of a concentrate, liquid, soft gel, driedpowder, capsule, pellet, pill, suspensions, emulsions, and a foodsupplement including health bars.
 5. The composition of claim 4 whereinthe product comprises between about 15% and about 25% by weight of themedicament and between about 65% and about 85% by weight of a filler. 6.The composition of claim 4 wherein the product comprises: a. betweenabout 15% and about 25% by weight of the medicament; b. between about60% and about 84% by weight of a filler; and c. between about 1% andabout 25% by weight of a dietary supplemental nutrient.
 7. Thecomposition of claim 1 in which the lignans are present in an amountfrom about 1% to about 70% by weight.
 8. The composition of claim 1wherein said therapeutic amount of said medicament is selected on abasis of a treatment for a coronary heart disease.
 9. The composition ofclaim 1 wherein said therapeutic amount of said medicament is selectedon a basis of a treatment for modulating a cardiovascular development.10. The composition of claim 1 wherein said therapeutic amount of saidmedicament is selected on a basis of a treatment for modulating bloodlipid profiles.
 11. The composition of claim 1 wherein said therapeuticamount of said medicament is selected on a basis of a treatment ofvascular conditions.
 12. The composition of claim 1 wherein saidisoflavones are selected from a group consisting of malonyl, acetyl,glucoside, and aglycone.
 13. The composition of claim 1 wherein saidcomposition is a food in a form of a concentrated, easy to consumedosage.
 14. A method of treating a human subject for a cardiovascularcondition, said method comprising the step of administering to the humansubject a therapeutically effective amount of a composition for treatingsaid condition, said composition being extracted from plant matter whichis enriched in at least two phytochemical fractions.
 15. The method ofclaim 14 wherein said phytochemical fractions are selected from thegroup of isoflavones, lignans, saponins, sapogenins, catechins, andphenolic acids.
 16. The method of claim 15 wherein said isoflavones areselected from a group consisting of malonyl, acetyl, glucoside, andaglycone and said composition is in a form selected from a groupconsisting of a concentrate, liquid, soft gel, dried powder, capsule,pill, suspension, emulsions, and food supplements including health bars.